Process to elaborate a biostimulant based on seaweeds

ABSTRACT

Process to elaborate a biostimulant based on seaweeds, comprising acid and alkaline treatment of the algae. The process comprises incorporating American Leonardite and subsequent spraying of product. The invention comprises the composition obtained and its use as germination promoter, root stimulator, among others.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of co-pending U.S. patentapplication Ser. No. 14/532,422, filed 4 Nov. 2014, which in turn claimsthe benefit of Chilean Patent Application No. CL 1464-2014, filed 4 Jun.2014, each of which is hereby incorporated herein as though fully setforth.

FIELD OF THE INVENTION

The present invention is related to a product based in seaweeds, that isuseful to be applied in all types of crops. More specifically, it refersto a biostimulant from seaweeds, that is useful to be applied in alltype of vegetal crops, either agricultural or forest, with the purposeto avoid chemical fertilizers that are polluting or harmful to humanhealth and to the environment.

BACKGROUND

Currently there is a wide range of methodologies to obtain seaweed-basedfertilizers.

The pending patent request WO2013/108188 describes a method to obtainfertilizers. The method the application of high pressures to seaweeds torelease its intracellular content. This extract is complemented withzeolite and meat and bone meal (MBM). The application does not claim theproduct in its current commercial form, i.e. pellet or grind down. Thisdocument is part of the state of the art.

Patent EP1534757 claims a process to obtain a fertilizer andphycocolloids in parallel. The extraction of the fertilizer includeswashing the seaweeds; blend the seaweed through milling; filtering andrecovery of the liquid phase; addition of a certain preservative;concentration through evaporation or by the use of membranes. The methoddoes not mention acid and/or base treatment, and for that reason theincorporation of a preservative is probably required. This patent ispart of the state of the art.

Patent CL 47236 from the Universidad de los Lagos claims a method thatincludes the use of HCl as the acidfying agent and K₂CO₃ as thealkalizing agent for the treatment of the seaweeds Macrocystis pyriferaand Ulva rigida, obtaining a liquid final product (Makromix).

Comparatively, the product of the present invention is superior to otherproducts obtained by comparable procedures, because of the particularityof the process and the raw material used, allow to achieve a higherbioavailability of the nutrients contained in the seaweeds and this istraduced in better yields, both in the modalities of foliar (soil)application as in the phase of seed germination.

In the quest of better field results, the present invention differs inkey aspects from the process that delivers a superior product, like theuse of a different pool of seaweeds, the use of acetic acid and KOH,that finally result in higher process yield and quality of the finalproduct.

DESCRIPTION OF THE INVENTION

Because of the growing demand on products that are organic,environmentally friendly and harmless to human health, the need fornatural biostimulants has been raised, that are similar or moreeffective that the traditional stimulants used. The present inventiondiscloses a biostimulant that is capable to increase the growth ratesand yields of a wide range of crops.

In order to efficiently obtain the product, with a better performance tothat described in the state of the art, the present invention claims amethod for the preparation of a natural vegetal biostimulant, thatincludes the treatments of seaweeds with acetic acid as acidifyingagent, and KOH as alkalizing agent. The present invention also includesthe use of american leonardite, to finally obtain a dry product, that iseasy to use, transport and store.

The present invention claims a method that takes advantage in theavailability of seaweeds of low commercial price as raw material toproduce a biostimulant, that is useful to be applied on seeds, aerialparts of the plants or soil. To accomplish this, red and brown algaefrom the Macrocystis and Gracilaria genera have been selected, like, forexample, M. pyrifera and G. chilensys, respectively.

Red and brown algae contain high concentrations of vegetal hormones orphytohormones, free amino acids and oligosaccharides. The compositionclaimed, due to the utilization of these seaweeds and the innovativemethod of elaboration, extracts and preserves, and in consequenceprovides to the vegetal or seed treated high concentrations of threephytohormones: auxins, cytokinins and gibberellins, that are requiredfor the optimal development of plants.

Furthermore, 17 amino acids present in seaweeds that are necessary forprotein synthesis (alanine, arginine, aspartic acid, cysteine, glycine,glutamic acid, histidine, isoleucine, leucine, lysine, methionine,phenylalanine, proline, serine, threonine, tyrosine and valine) ensure abetter development of the plants. In normal conditions plants producetheir own amino acids, but in stress conditions photosynthesis isreduced and in consequence its metabolism is slowed down. The productfrom this patent, delivers exogenously these amino acids and reduces thestress in plants, increasing the absorption of nutrients and improvingits translocation and the permeability of the cell membranes.

In addition, oligosaccharides (laminaran, manitol) activate the functionof phytoalexins in cultivated plants, resulting in a higher vigor andresistance of the plant to diseases and plagues, optimizing thetechnical use of them.

In sum, the method claimed allows the extraction of nutrients containedin seaweeds foliage, without degradation nor requiring thereincorporation of those nutrients from external sources.

Finally, the present invention provides a great amount of organicmatter, that is beneficial to improve the soil treated, by means ofhumic and fulvic acids incorporation that come from leonardite, that isa fossil mineral that is found together with lignite and that is themain source of these acids. This solid, concentrated and humidifiedorganic matter, contain humic substances, that help to improve the soilsboth in a physical-chemical and biological manner. It can be appliedtogether with solid and liquid fertilizers to obtain a betterperformance. Is an intermediate material, between turf and lignite, andis derived from the transformations (diagenesis) of vegetal remains,buried at approximately 10 meters, when together with water percolationfrom rain and the presence of atmospheric oxygen, led to a progressiveenrichment of the humic substances.

It is possible that brown lignite may be confused with leonardite, eventhat they have clear differences: lignite suffers a carbonizationprocess, whereas leonardite does not, and at the same time, leonarditehas a high oxygen content and lignite loses almost all of it, andfinally, leonardite has an open structure, and lignite comprises,because of the pressure suffered during its burial.

Main Functions of Leonardite:

Acts as a soil corrective, achieving the rehabilitation of soils byproviding them a fluffy structure, reducing compression and favoringventilation and porosity of soil; it helps to water retention; it favorsroot growth; it reduces the need of mineral fertilizers; it improves thequality and size of crops; it accelerates the plant's vegetative cycle;it improves the soil physical-chemical properties; it improves the soilsalinity; it has a high power to chelate cations.

FIGURE DESCRIPTION

FIG. 1: Flowchart that exemplifies the complete process for biostimulantextraction.

DETAILED DESCRIPTION OF THE INVENTION

In the following pages the physical-chemical fundaments are describedfor each of the steps of the process and the evidence that show theoptimal results as well as the detailed procedure to reproduce thoseresults with the available resources.

Seaweed Milling

Seaweed milling has as primary aim to facilitate the use of them duringstorage, salt removal or washout and the acid treatment. Another benefitderived from milling is to reach a particle size that allows to reagentsuniformly penetrate to the seaweed, resulting in more homogenousreactions and so it gives better monitoring of chemical treatments. Theuse of this maneuver will result in obtaining fragments, that makepossible to perform in a satisfactory way the subsequent treatments.This is done using a grinder. With this mechanism a more uniformparticle size is obtained resulting in more mass fluidity duringseaweed's chemical treatments. The preferred particle size rangesbetween 5 and 10 mm.

For processing in the factory, 1000 kg of fresh seaweed are used, theyare deposited in a 5000 L capacity vat (tank). If seaweeds will not beimmediately processed, they can be stored for 24 hours withoutfreshwater.

Seaweeds are employed in a ratio of Macrocystis:Gracilaria from between70:30 to 95:5 respectively.

Salt Removal or Washout

This step consists in washing the seaweeds with water, in order toeliminate and remove mineral salts, seawater excess, sand, smallmollusks, etc. The process is carried out with water at room temperaturefor until 60 min.

Acid Treatment of Seaweeds

The acid treatment step has two basic functions. First it is done toremove the soluble mineral salts and organic matter in excess, that wasnot removed by the previous wash, as well as the sediments and organismsassociated to seaweeds. Second, is to perform an ion-exchange chemicalreaction mainly between calcium ions and other divalent cations, likemagnesium and strontium, that are contained in seaweeds as divalentmetal alginates, and originating alginic acid (HAlg).

Although there is currently in the state of the art a process thatemploys HCl, we have discovered that the use acetic acid results insuperior acid treatment. Both HCl and acetic acid run in a similar wayin the chelate fixation that is required in the acid process. However,acetic acid is accepted in the organic process without HCl limitations,such as its low boiling point, that implies a higher acid loose at thetemperature in which the acid treatment is done, with respect to aceticacid. Additionally, HCl treatment only tolerates a 1.5% of the acid, anamount that is not enough for an efficient chelate fixation in a shorttime of reaction.

1. The Reaction Carried Out is the Following:

Ca(Alg)₂+2CH₃COOH----->2HAlg+Ca(CH₃COO)₂

Physically, this is a heterogeneous reaction between solid seaweedparticles and the acid solution.

2. Liquid/Solid (L/S) Relation.

The ratio of water respect to the volume of processed seaweeds has twopurposes; first, is to provide fluidity to the mass in order to obtainan homogeneous reaction, second is to remove the mineral salts andsolids associated to seaweed particles.

a) Factors Implied in Mass Fluidity.

Mass fluidity depends not only in the liquid amount but also in theparticle size. The smaller the size of the particle the lesser theamount of water required. However, the minimal size of the particle islimited by two important factors:

-   -   Viscous nature of seaweeds: Naturally, fresh seaweed have a        viscous texture, which is incremented after fractioning by the        release of fucoidins in the cutting edges.    -   Separation of liquid: solid phases: Very small particles are not        convenient because of the complications in the mechanisms to        separate solids and because of the increased loss of small        particles

In conclusion, a particle size between 5 to 10 mm present an homogeneousfluidity with a minimal liquid solid relationship of 2:1.

b) Removal of Mineral Salts and Soluble Organic Matter.

Humid seaweeds contain an 8-10% of mineral salts and soluble material.These impurities often causes interferences in some of the steps of theprocess of preparation of the biostimulant, so it is necessary to reducethem at the beginning of the process. A 1.5-3.0% reduction avoidsignificant interferences in subsequent reactions. In previousexperiences, the process for salt removal was standardized to 1.0-3.0%levels, applied a 30 minutes wash in a liquid:solid relation of 2:1.With this washing system the water consumption is optimized and anefficient reduction in salt content is achieved.

3. Acid Consumption.

During the acid wash of seaweeds a ion-exchange reactions occurs betweende divalent metal salts from alginic acid that is contained in seaweeds(Ca²⁺, Mg²⁺, Sr²⁺) and proton H⁺. This treatment is done in order totransform all the alginate salts contained in the seaweed to its genericform of alginic acid.

Depending on the alginate content in the seaweed, this reaction consumesan amount of acid close to the stoichiometric relationship.

In practice, the total expense of acid, including the lost in residuesshould be the closest to said relationship. This will be depend on theefficiency of the wash mechanism.

The washing system suggested for salt removal was used to optimize acidconsumption.

The average expense recorded is between 55 and 65 L of acetic acid, morepreferentially between 58 and 62 L of acetic acid for 1,000 kg of freshseaweed.

4. Time and Temperature of Reaction.

Time and temperature notably influence a reaction's kinetics. Theseparameters determine the viscosity of the final product due to thesusceptibility of alginic acid to break down at high temperatures orprolonged exposures to acid.

The reaction time is from 90 to 120 minutes at a temperature of 45-65°C., time that is satisfactory to complete the reaction y there is noalginate degradation, protecting the compounds from seaweeds, andobtaining a chelated base.

The parameters previously mentioned are resumed in Table 1:

liquid/solid relationship 2:1 Amount of acetic acid 55 to 65 L Totaltime of residency 90-120 min Temperature of reaction 45-65° C. Wash 30min Batch Size 1,000 kg of fresh seaweeds

Once the acid treatment is finished, the mixture is filtered with sieve,recovering the solid particulate, that will be subject to digestion oralkaline treatment.

Digestion and Extraction (Alkaline Treatment)

During digestion and extraction a neutralization reaction occurs betweenalginic acid contained in algae particles and a potassium alkali,producing in this case a potassium alginate en aqueous solution. Thereaction carried out is the following:

HAIg+KOH------------>KAlg+H₂O

This reaction is one of the most delicate of the process, because a bigproportion of the performance and the quality of the final productdepends on the adequate control of the physical-chemical parameters thataffects it.

Physically, this is a heterogeneous reaction between the solid particlesfrom the acid pretreated seaweed and the alkaline solution. The alkalipenetrates in the algae particle, converting the insoluble alginic acidin soluble sodium or potassium alginate in aqueous media, finallyresulting in a very viscous solution with millimetric insolublecellulose filaments. The total dissolution of the particles determinethe maximal yield of the raw material.

Basically the reaction is completed at a pH higher than 7,preferentially 8.5 to 14, in a reaction time that can range from 120 to150 min depending on the following factors:

a) Temperature. The temperature is one of the most important factors inthe reaction due to its control in the reaction's kinetics and theviscosity of the final product. The range in which the viscosity can becontrolled varies from 70 to 82° C.

b) Particle size: A particle's size <10 mm gives very good results withthe appropriate combination of the other parameters.

c) liquid:solid relationship and stir. During digestion is recommendedto have a fluidity that allows to homogeneously stir the suspensionusing minimal water.

A 2:1 liquid:solid relationship respect to fresh seaweeds results in aviscosity that allows a homogenous stir of the suspension and givesenough aqueous media to dilute the extract.

d) pH. In the process described optimal results were obtained using a pHrange between 8.5 and 14 (given by the use of potassium hydroxide).Under these conditions maximal yields between 3.5 and 3.8% of potassiumalginate were obtained, that are the base of the alkaline extract usedas biostimulant from fresh seaweeds. The alkali consumption is between45 and 50 kg of potassium hydroxide per initial ton of fresh seaweed.

The parameters previously mentioned are resumed in Table 2:

Temperature 70-82° C. pH 8.5-14 Time 120-150 min liquid/solidrelationship 2:1 % of KOH respect to fresh seaweeds 3 to 6%The digestion is carried out as follows:

Potassium hydroxide is dissolved in water at 60-65° C. and milledseaweeds or the solid particulate recovered from the acid treatment areadded, then the mixture is stirred for 120 to 150 min at 70-82° C. pH isadjusted with potassium hydroxide. The final product of this reaction isa viscous solution (400-600 Cps) of potassium alginate with millimetriccellulose residues.

The end of the hydrolysis step is initiated lowering the temperature ofthe mixture, followed by the filtration of the alkaline extract.

5. Filtration

The aim of filtration is to clarify the solution of the alkalineextract, removing the cellulose insoluble particles that remains fromdigestion step. The monitored parameters in this stage are the filtratepurity and filtrate velocity, which depends on the filtration mediaused. Preferentially the filtration of the solution is carried out witha primary and a secondary filter with a final net size of 50, withoutexcluding another compatible methods.

6. Leonardite Processing

Leonardite is subjected to an activation process through chemicalhydrolysis to separate it in humic and fulvic acids (active components)and other non hydrolysable components (clays and humines). This helps toextract all the nutrient capacity of leonardite in short time, processthat in a natural manner will take several years.

Digestion is done in aqueous media, with KOH and controlled temperature.For each 100 kg of leonardite 500 L of water at 40-45° C. are used, andbetween 47 and 53 kg, more preferentially between 50 and 51 kg of KOH.The mixture is stirred at 45-50° t for 30 to 40 min.

Once the digestion of leonardite is finished, it is mixed with thealkaline extract from seaweed, recovered from the alkaline treatment, ina concentration that ranges from 5 to 20% v/v.

7. Drying

The process consists in pulverize the fluid inside a chamber subjectedto a controlled stream of hot air. This fluid is atomized in millions ofindividual microdrops by a rotating disc or a spray nozzle. By thisprocess the area of contact of the pulverized product is enormouslyaugmented and inside the chamber the hot air stream induces the rapidvaporization of water in the center of each microdrop were the solid islocated, resulting in a smooth dry without a big termal shock,trasforming the product in powder and finishing the process with thecollection of this powder.

The final product is a fine powder, resulting from the drying of themixture from the extract from processed seaweeds and the solubilizedamerican leonardite. This presentation facilitates transportation,storage and preservation, keeping the biological properties of theproduct.

For application on ground, the product is simply dissolved in water,reconstituting the extract (28 gr×L) depending on the requirements,being able to be applied by irrigation or spray.

The product has been stored under these conditions for periods longerthan 3 years, keeping the biostimulant properties of the product, afeature that is favored for its powder presentation format and its lowhygroscopy.

EXAMPLES Monitoring of a Consignment in the Factory.

The following is a description of a typical operational process in thefactory including the monitoring that is performed in every step of theprocess starting with a base of a 1,000 kg consignment of freshseaweeds.

Preparation of a Consignment

Seaweeds must be fresh. At the beginning of the process the followingpreparations must be done:

2,000 L of water with 61 L of acetic acid are prepared in the acid washtank.

2,000 L of water are stored in the digestion tank.

Reagents are heated to a temperature close to 45-50° C. and the acidwash can now begin.

Procedure of the Operation

1. Collect 900 kg of Macrocystis pyrifera and 100 kg of Gracilariachilensis.2. Fraction them to a size between 5 to 10 mm in the milling machine,wash and weight 1,000 kg;3. Incorporate 2,000 L of water and 61 L of acetic acid at 45-65° C.with constant stirring for 100 min. Once finished, the liquid phase iseliminated and it is washed with water.4. 2,000 L of water are added, and the temperature is set to 60° C.; 48kg of potassium hydroxide are dissolved and milled seaweeds are added.Temperature is raised to 79° C. under constant stirring for 2 hours. pHis adjusted at 8.65 and the mixture is cooled down.5. Filtration. Once the residual particles have sedimented, a filtrationtakes place, eliminating the particulate; american leonardite isincorporated, hydrolized at 10% v/v.6. Adjust pH to 7, if required, with phosphoric acid, acetic acid orother suitable acids.7. Dry until obtain the powder.

6. Leonardite Processing For Obtaining Humic and Fulvic Acids.

500 L of water at 45° C. are employed. The mixture is stirred until theend of the process. 50 kg of KOH and 100 kg of american leonardite areadded. Temperature is hold at 45° C. for 30 to 35 minutes, until theinitial product is completely dissolved.

Example 1: Comparative Assay in Wheat

Variety Pandora INIA Seed dose 220 kg × Ha Total area 18 haBioestimulant application area  1 ha Makromix application area  1 haControl area 16 ha

Treatment was applied to seeds pre-sowing and then to the plant in theend of culm state.

SCHEDULE OF APPLICATIONS Date/period Product Dose Watering Area 15-Jul/Biostimulant 0.5 L × 100 kg of seed 1 L × 100 kg  1 ha Pre-sowing15-Jul/ Makromix 0.5 L × 100 kg of seed 1 L × 100 kg  1 ha Pre-sowing25-Oct/ Biostimulant 3 L × ha 66 L × ha 16 ha End of culm 25-Oct/Makromix 3 L × ha 66 L × ha 16 ha End of culm

Both treatments showed differences respect to control, in the measuresof root growth, vegetative development and plat height, evidencingnotorious differences in harvest yield and the milling quality.

MEASUREMENTS 05-Nov Biostimulant Makromix Control Root growth 15 14.5 14Roots lenght m/m 40 38 38 Plant weight Grs. 91 78 68 HARVEST 15-FebBiostimulant Makromix Control qq × ha. 71.3 66.23 60.46 CALIDAD MOLINERA20-Feb Biostimulant Makromix Control Hectoliter weight kg/HL 85.3 85.384.6 Dry gluten kg/HL 11.93 10.4 9.9

Example 2: Comparative Assay in Raspberry

In raspberry crop, Meeker variety, the following assay was performed ina total area of 1.98 ha:

Bioestimulant application area 0.92 ha Makromix application area 0.94 haControl area 0.12 ha

Four applications of each fertilizer were done, at a rate of 6.5 L perha, in a period of 80 days. In both applications a higher development ofbuds and shoots as well as an apparent increase in vigor, additionally ahigher need of fertilization is observed, mostly nitrogen after eachapplication, a fact that is probably explained by the increase in theplant's rate of biomass development.

The yield in kg of pulp per hectare was evaluated. Yield of eachtreatment according to quality and expressed in kg/ha.

Classification According quality Biostimulant Makromix Control IQF > 182,777.75 2,509.66 2,430.48 IQF < 18 1,860.38 1,869.00 1,789.37 Pulp486.875 438.347 463.1487 Total yield 5,125 4,817 4,683

Example 3: Comparative Assay in Eucalyptus

A seed from Puacho 3 (Anchile) was employed, subjected to 3 days ofsoaking in water and 30 seconds of soaking in Sodium hypochlorite, 21days of cold and 3 days in chamber. Pine bark from Madexpo and 3 kg ofNPK (3-33-3) was used as substrate.

30 seed trays with 96 cavities each were used. A solution of 1% ofBiostimulant and Makromix was applied. A second application wasperformed at a concentration of 0.5%.

Procedure:

Two treatments plus the control (Ultrasol) were done, each one with 5replicas. Sowing was carried out using an automatic sow seed machine.After that a solution with 1% fertilizer was applied with a wateringcan. Then, trays were taken to the germination chamber at a fixedtemperature of 27° C. for 3 days.

Once initiated germination, trays were taken to a farm building withtemperatures ranging between 20 and 30° C. during day and 4 and 15° C.during night. Watering, depending on weather conditions, was done twicea day.

The controlled variables controlled were the germination percentage,root collar diameter (RCD) and average height after 10 days of sow.

Second application was done with 0.5% dilution in 1,500 cc of water.After 36 days of sow, a second control of germination was done.

Results were the following:

% Germination Average height 1st 2nd 1st 2nd Fertilizer measurementmeasurement measurement measurement Biostimulant 77.5 77.08 1.08 2.74Makromix 59.79 58.54 1.02 2.12 Ultrasol 76.88 75.83 1.04 2.70

Example 4: Comparative Assay in Grape

Variety Red Globe Location Copiapó - Chile Cultivated field 8 yearsTotal area 5 ha Biostimulant applied area 2 ha Makromix applied area 2ha Control area 1 haThe following application protocol was employed for both products:

Application period Dose Watering 40 cm bud 3 L × ha 1.6 L × ha Curdle 3L × ha 1.6 L × ha Bunch closure 3 L × ha 1.6 L × ha Veraison 3 L × ha1.6 L × haParameters were measured 15 days pre-harvest and during harvest.

Biostimulant Makromix Control measurement 15 days pre-harvest Bunchweight 506 490 456 Equatorial diameter 19.5 19.2 19 Brix degrees 18.618.4 18.3 Harvest Bunch weight 694 642 636 Equatorial diameter 23.8 22.121.96 Polar diameter 2.8 2.7 2.7 Brix degrees 19.35 19.1 19

It is evident the differences between treatments and control, in all theparameters measured, being the treatment with Biostimulant notoriouslysuperior with respect to Makromix and Control.

Example 5: Comparative Assay in Tomato

Variety Titan Total Area 0.6 ha Biostimulant Area 0.3 ha Makromix Area0.3 ha Layout of plantation 1.2 m between row y 0.2 over row WateringDrip irrigation with tape. 4 L × m linear Applications 8 foliarapplications with 400 L × ha Dose 2 L × ha in C7

Eight applications of each treatment were done, every 20 days. In the 2first applications no differences were observed in plant development,however, starting the third application the vigor of theBiostimulant-treated seedlings was increased, resulting in a notoriousincrease in total yield and in a better fruit quality at harvest.

HARVEST Kg × ha Quality Bioestimulant Makromix Tomato 1^(st) 175,109117,600 Tomato 2^(nd) 50,031 39,210 Tomato 3^(rd) 25,015 39,190 Total250,155 196,000

What is claimed is:
 1. A method of producing a biostimulant fromseaweed, the method comprising: a) milling fresh seaweed; b) washing thefresh seaweed c) treating the washed seaweed with an acid; d) treatingthe product of c) with KOH; e) separating and recovering the liquidphase of the product of d); f) adding hydrolized American leonardite tothe liquid phase; and g) drying the product of f) to obtain a powder. 2.The method of claim 1, wherein the seaweed is selected from a groupconsisting of Macrocystis spp. and Gracilaria spp.
 3. The method ofclaim 1, further comprising: milling the seaweed to a particle size ofbetween about 5 mm and about 10 mm.
 4. The method of claim 1, whereinwashing the seaweed is carried out with water at room temperature. 5.The method of claim 1, wherein the acid includes acetic acid.
 6. Themethod of claim 5, wherein the acetic acid is employed at a ratio ofbetween 55 L and 65 L per 1,000 kg of seaweed.
 7. The method of claim 1,wherein treating the washed seaweed with an acid is carried out atbetween 45° C. and 65° C. for between 90 minutes and 120 minutes.
 8. Themethod of claim 1, wherein treating the product of c) with KOH employsKOH at a ratio of between 45 kg and 50 kg per 1,000 kg of seaweed. 9.The method of claim 1, wherein treating the product of c) with KOH iscarried out at between 70° C. and 82° C. for between 120 minutes and 150minutes.
 10. The method of claim 1, wherein treating the product of c)with KOH is carried out at a pH between about 8.5 and about
 14. 11. Themethod of claim 1, wherein treating the product of c) with KOH includescooling the product of c).
 12. The method of claim 1, wherein separatingincludes filtering the product of d) with a primary and a secondaryfilter with a final net size of
 50. 13. The method of claim 1, whereinadding hydrolized American leonardite includes adding hydrolizedAmerican leonardite at a range between 5% and 20% v/v.
 14. The method ofclaim 1, wherein the hydrolized American leonardite is hydrolyzed byalkaline treatment with between 47 kg and 53 kg of KOH per each 100 kgof American leonardite at between 40° C. and 50° C. for between 30minutes and 40 minutes with constant stirring.
 15. The method of claim1, wherein drying the product of f) includes spray drying.
 16. Abiostimulant composition prepared according to the method of claim 1.17. The biostimulant composition of claim 16, wherein the composition issuitable for storage for at least three years in the form of a drypowder.
 18. A rehydrated biostimulant composition prepared according tothe method of claim
 1. 19. A method of improving plant growth, themethod comprising application of a biostimulant according to claim 18 toat least one of soil, a plant, or a seed.
 20. The method of claim 19,wherein improving plant growth includes at least one of the following:promoting seed germination, stimulating root development, improvingresistance to extreme weather conditions, including drought and freeze,increasing resistance to disease, increasing resistance to insectattack, prolonging a vegetative period, increasing a period ofproduction, or increasing a period of harvest.